tbuckley/cabot-search huggingface dataset:
Abstract P3130: Role Of The Placenta-derived Extracellular Vesicle-cargo Protein, Vasorin In Vascular Dysfunction In Preeclampsia
Authors: Saravanakumar Murugesan, Mark F Powell, Hanna Hussey, Lakshmi Saravanakumar, Adam B Sturdivant, Rachel G Sinkey, James A Mobley, Tamas Jilling, Dan E Berkowitz
Journal: Circulation Research (2022)

Background: We recently demonstrated that placenta-derived extracellular vesicles (pEVs) cargo play a role in vascular dysfunction in patients with severe preeclampsia (sPE). We focused our follow up studies on a novel protein vasorin (VASN) that has not as yet been studied in PE. Here, we hypothesize that decreased VASN in EVs cargo plays a significant role in the regulation of vascular endothelial proliferation and function in sPE. Methods: We manipulated VASN by overexpression and knockdown in human aortic endothelial cells (HAEC) and in murine aortic vascular rings (MVR) using adenoviral vectors encoding VASN (AD-VASN) or VASN shRNA (AD-shVASN), respectively. VASN levels in EVs, in HAEC and in MVR with/wo adenoviral overexpression or knockdown of VASN was done by western blotting. Migration of HAEC was studied by a scratch and repair migration assay and contractile function of MVR was assessed by wire myography. A PE-like vascular dysfunction was produced in timed pregnant mice by intravenous injection of an adenoviral vector encoding the short form of FMS-like tyrosine kinase 1 (sFLT-1) on embryonic day 10.5 (E10.5) of pregnancy. Plasma from sFLT-1 -injected and control untreated (UT) mice were collected on E15.5,E17.5 and E19.5. Results: We confirmed our findings with proteomics and using western blotting we detected a significant reduction in VASN levels in both urinary and plasma-derived EVs in sPE patients when compared to normotensive. Moreover, we detected a time-dependent significant reduction of VASN in EVs from plasma of sFLT-1 injected mice whereas UT mice exhibited a time-dependent increase of VASN. Western blotting also confirmed over-expression and knock down of VASN in HAEC and in MVR treated with AD-VASN and AD-shVASN, respectively. Treatment of HAEC with EVs from sPE, but not by EVs from NTP inhibited migration, and pretreatment with AD-VASN but not with AD-shVASN rescued migration upon treatment with EVs from PE. Treatment of MVR with EVs from sPE, but not by EVs from NTP inhibited acetylcholine (ACh)-induced vasorelaxation. Pretreatment with AD-VASN, but not with AD-shVASN rescued ACh-induced vasorelaxation upon treatment with EVs from sPE. Conclusion: Over-expression of VASN using adenoviral vectors rescued inhibitory effects of EVs from sPE on migration of HAEC and ACh-induced vasorelaxation in MVR, suggesting that reduced VASN content in EVs from sPE plays a mechanistic role in vascular dysfunction observed in sPE.
Abstract P2106: Arrhythmia As Cause For The Development Of The Takotsubo Syndrome In A Patient-specific Atrial Stem Cell Model
Authors: Charlotte Brandt, Daniela Huebscher, Gideon Syed-Ali, Viacheslav O Nikolaev, Katrin Streckfuss-Boemeke
Journal: Circulation Research (2022)

Background and Aims: The Takotsubo syndrome (TTS) is characterised by an acute left ventricular dysfunction without exhibiting signs of stenosis. TTS affects mainly the left ventricle, however, its pathophysiology comprises transient impairments in left atrium functions with a prevalence of atrial fibrillation (AF) of around 18% correlating with higher ventricular heart rhythm disorders. In this study, we aimed to prove the hypothesis that molecular and cellular arrhythmic events contribute to the development of TTS under catecholamine stress and to test if the currently clinically used β-blockers (Metoprolol) are suitable for Takotsubo cardiomyopathy in vitro. Methods and Results: We generated induced pluripotent stem cell-derived atrial cardiomyocytes (TTS-iPSC- a CMs) from TTS patients, confirmed atrial marker expression (MLC2a, PItX2, NR2F2), and treated the cells with catecholamines to mimic TTS-phenotype. Using a cytosolic Förster resonance energy transfer (FRET) based cAMP sensor, we tested the activity of phosphodiesterases (PDEs) in TTS-iPSC-aCMs, which are besides β-ARs main players involved in cAMP signaling in CMs. We observed that after β-AR stimulation, the strong effects of the PDE4 family in the cytosol of atrial controls were significantly decreased in aCMs of the TTS patients. This effect was completely rescued after application of PDE4 activator MR-L2 and is in line with the previously described downregulation of PDE4 in human atrial myocardium of AF patients. In functional studies, isoprenaline-induced increase in systolic Ca 2+ transient amplitude was more pronounced in TTS iPSC-aCM compared to controls. These effects were rescued by both, the clinically approved β-blocker Metoprolol as well as the PDE4 activator MR-L2. To analyse arrhythmic events in atrial TTS CMs, we performed confocal Ca 2+ measurements and demonstrated that the diastolic sarcoplasmic reticulum Ca 2+ leak was increased in iPSC aCMs of TTS patients compared to control under basal conditions and after Iso-treatment. In addition, TTS patients displayed faster Ca 2+ kinetics compared to control cells, already under basal conditions. These results were underlined on a molecular level by increased phospholamban phosphorylation in TTS iPSC-aCM .Subsequent treatment with the beta-blocker Metoprolol rescued the Ca 2+ kinetic parameters in all cell lines. Conclusion: In conclusion, we were able to draw a comprehensive picture on the role of the arrhythmia in the development of TTS. We found TTS-specific differences with reduced PDE4 activity, elevated arrhythmic events and enhanced reactions to catecholamines, which could be rescued by the clinically approved drug Metoprolol and partly by the PDE4 activator MR-L2. Therefore, Metoprolol as well as MR-L2 demonstrate promising effects as patient-specific therapeutic target for TTS.
Abstract P3133: Endothelial Nup93 Selectively Regulates The LaminB1 Isoform To Limit Features Of Cellular Senescence And Consequent Endothelial Dysfunction
Authors: Tung D Nguyen, Mihir K Rao, Mary G Horne, Michael A Winek, Monica Y Lee
Journal: Circulation Research (2022)

Recent translational studies have focused on the role of the nuclear pore complexes (NPCs) and its component nucleoporin proteins ( i.e. nucleoporin93 [Nup93]) as potential mediators of cardiovascular disease (CVD). The nuclear pore complex (NPC) is one of the largest structures found in the nuclear envelope, serving as the main transport conduit between the nucleus and cytoplasm. While historically perceived as static gateways with the sole purpose of nucleocytoplasmic exchange, recent investigations implicate nucleoporins in several transport-independent features ( e.g. nuclear lamin organization). Here, we show that endothelial loss of Nup93, a critical NPC protein, selectively reduces expression of the laminB1 isoform while enhancing γ-H2AX-Ser139 phosphorylation, both well-known markers for cellular senescence. Co-immunoprecipitation experiments in Nup93-enriched fractions indicate a preferential association with the LaminB1 isoform, when compared to Lamin A/C. Furthermore, endothelial depletion of Nup93 promotes inflammation, increases stress fiber formation, and enhances paracellular permeability - all features of cellular senescence and ensuing EC dysfunction. Collectively, our data demonstrate that a Nup93-LaminB1 interaction may be required to maintain adequate LaminB1 expression in ECs, where the age-associated decline in Nup93 levels may selectively reduce LaminB1 isoform levels to drive the consequent pathological features of endothelial senescence, a known contributor to vascular aging and CVD.
Abstract P2081: Beclin1 Ameliorates The HIV-1 Induced Cardiac Dysfunction In Mice Model Of HIV
Authors: Shiridhar Kashyap, Maryam Rabbani, Isabela Beatriz Futuro de Lima, Sameep Gawasker, Raj Patel, Jay Rappaport, Qiangrong Liang, Manish K Gupta
Journal: Circulation Research (2022)

Background: Despite antiretroviral drugs-mediated control of active HIV infection, cardiovascular disease (CVD) remains one of the leading causes of comorbidity in people living with HIV/AIDS (PLWHA). Recent studies suggest that activation of the immune system and chronic inflammation increases CVD risk in PLWHA. Dysregulation of the immune system can interact with several cellular functions, including autophagy and mitophagy. The autophagic factor Beclin1 is a crucial regulator of cellular autophagy and mitophagy. In this study, we tested the role of Beclin1 in cardiac function using a mice model of HIV. Methods and Results: This study was performed using transgenic mice of HIV-1 Tg26 mice, which was constructed before using HIV-1 DNA with deletion of 3-kb region of the gag/pol genes. Tg26 mice were crossed with Beclin1 transgenic mice to test the role of Beclin1. Cardiac function of Tg26 and Tg26 mice crossed with Beclin1 was evaluated by echocardiography. Cardiac autophagy was determined by western blots using autophagy regulatory markers. Our data suggest that Tg26 mice have significantly reduced cardiac function. Further, Tg26 mice show increased reactive Oxygen species (ROS) level, accumulation of LC3II, increased P62 level, and reduced autophagy flux. Additionally, western blots with cardiac tissue show upregulation of electron transport chain (ETC) complex proteins of mitochondria in the Tg26 mice that may be due to defective mitochondria accumulation in Tg26 mice heart. Interestingly, Beclin1 expression can improve the cardiac function of Tg26 mice heart. Mechanistically, we found that Beclin1 overexpression can restore the autophagy flux and reduce the cellular ROS level in the heart of Tg26 mice. Conclusion: Our study suggests that Beclin1 plays critical role in regulation of cardiac function during HIV induced cardiomyopathy and can be used as a potential therapeutic tool to control comorbidity in the PLWHA.
Abstract P3028: Animal In Vivo: The Proof Of Flow Choking And Bulging Of The Downstream Region Of The Stenosis Artery Due To Air Embolism
Authors: Anbu J, Deveswaran R, Bharath S, Sanal Kumar V R, Shiv Kumar Choudhary, Pradeep Kumar Radhakrishnan, Bharath Rajaghatta Sundararam, Charlie Oommen
Journal: Circulation Research (2022)

Background: Evidences are escalating on the asymptomatic cardiovascular diseases and disorders associated with air embolism due to the flow choking (biofluid/Sanal flow choking) [PMID: 32837737 &amp; PMID: 34326352]. Flow choking occurs anywhere in the circulatory system at a critical systolic to diastolic blood pressure ratio (BPR) &lt;https://doi.org/10.1161/res.129.suppl_1.P422&gt;. Flow choking leads to the shock-wave generation followed by pressure overshoots if the downstream region of the choked artery is having divergence/bifurcation regions. Methods: Animal in vivo study has been carried out on 6 th December 2021, IST: 2:00-4:00 PM (the FIRST in the World) for establishing the phenomenon of flow choking in a stenosis artery at a critical BPR due to air embolism. The high pressure air is injected into the upstream region of the stenosis using a syringe (see Fig.1(a)). Stenosis is created using a knot. Syringe infusion pump is used for altering the flow rate and upstream pressure. The multiple animal in vivo studies were carried out on 31 st December 2021 . It aims for demonstrating the phenomenon of flow choking followed by bulging and/or tearing of the downstream region of the stenosis artery of a small animal. Results: Figure 1(b) shows the animal in vivo (with the approval of the ethics committee) for the live demonstration of flow choking and bulging in a stenosis artery due to air embolism. Herein, we established the proof of the concept of flow choking due to air embolism in the stenosis-artery at a critical BPR (SBP/DBP) causing the risk of asymptomatic cardiovascular diseases and disorders. Conclusions: We concluded that at a critical BPR, the flow choking occurs in any artery. The shock wave generation due to flow choking leads to cardiovascular risk. Frequent flow choking creates memory effects (stroke history) in the downstream region due to the enhanced wall stiffness as a result of pressure-overshoot due to multiple shock waves. We further concluded that the critical BPR is a risk factor of flow choking causing asymptomatic cardiovascular disease, if the downstream region of the choked artery is having divergent shape.
Abstract P2103: Cannabidiol Protects Cardiac Function <i>In Vivo</i> And Inhibits Endothelial-to-mesenchymal Transition <i>In Vitro</i> : A Potential Anti-fibrotic Therapy For Heart Failure.
Authors: Muthu Kumar Krishnamoorthi, Forughalsadat Sanaee, Afsaneh Lavasanifar, Keith Youker, Arvind Bhimaraj
Journal: Circulation Research (2022)

Introduction: Cannabidiol (CBD) has been investigated for application in various diseases including cardiovascular ailments. Endothelial-to-Mesenchymal transition (EndoMT) has been reported to contribute to cardiac fibrosis and eventually heart failure (HF). Here we demonstrate that both free (f) and nanoparticle encapsulated (n) CBD preserves cardiac function in a non-ischemic mouse model of HF. Further, we show that CBD inhibits EndoMT in vitro . Hypothesis: CBD administration will improve cardiac function by targeting anti-fibrotic mechanisms. Methods: HF was induced in a 3-month old C57BL/6J mice (n=3) with 1% NaCl, 0.3 mg/mL L-NAME and an osmotic pump infusion of angiotensin II (Ang II). Both fCBD and nCBD were administered twice per week subcutaneously and echocardiography and histology evaluated at 5 weeks. We used an in vitro EndoMT model with L-NAME+ Ang II as EndoMT inducers in HUVECs. CBD at 0.1, 1 and 2 μM was used to study the dose impact. Results: HF mice treated with fCBD, and nCBD exhibited significantly higher EF when compared to untreated HF mice (Fig. 1 A). Histological evaluation showed that nCBD treated group displayed a higher anti-fibrotic activity compared to fCBD (both with less fibrosis compared to HF mice) (Fig. 1 B). In vitro , when CBD was added during EndoMT induction, the transition process was inhibited. Immunofluorescence studies exhibited a dose dependent effect of CBD in inhibiting Vimentin (mesenchymal marker) expression during the EndoMT process (Fig. 1 C). Conclusion: In conclusion, CBD protects cardiac function and exhibits an anti-fibrotic effect, possibly mediated by EndoMT inhibition. A nano formulation of CBD might have a more potent impact.
Abstract P1141: Heparinized Alginate And Collagen-based Hydrogels Enhance Localized Vascularization In Ischemic Tissue
Authors: Stephanie M Roser, Alicia Minor, Collin Polucha, Kareen L Coulombe
Journal: Circulation Research (2022)

Over 800,000 myocardial infarctions (MI) occur annually in the United States alone, 25% of which represent a repeat occurrence further indicating chronic ischemia in the myocardium. Cardiovascular regeneration is largely dependent on the stimulation of blood vessel development for recovering tissue perfusion and modulating ventricular remodeling. The delivery of angiogenic growth factors and cytokines has demonstrated the ability to initiate angiogenesis and vascular network formation in cardiac tissue, with localization of bioactive molecules more effective than systemic delivery. Alginate, with its high biocompatibility and biodegradation, represents a promising hydrogel material for factor delivery due to its high protein loading capacity. We hypothesize that covalently binding heparin to alginate will increase binding and retention to subsequently lengthen the local release duration of vascular-promoting factors. For greater tuning and biointegration, collagen and alginate hydrogel designs were optimized for both injection and co-implantation of a solid film with hiPSC-CM based ECTs in a rat model of myocardial infarction. Injectable and solid formulations showed stiffnesses of 50 +/- 2 kPa and 900 +/- 193 kPa, respectively, which was maintained for up to 6 hours of dry time. Release profiles showed sustained growth factor release, with 65% of the total content released at 7 days and 100% release in 14 days. In vitro evaluation using human induced pluripotent stem cell-derived endothelial cells (hiPSC-ECs) showed cell viability and primitive network formation. In vivo subcutaneous injections showed host-vessel penetration into the delivered biomaterial within 4 days. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) loaded injections demonstrated an average lumen density of 80 +/- 15 vessels/mm 2 when compared to 4 +/- 1 vessels/mm 2 in the unloaded controls. In conclusion, we have designed a biomaterial system for disparate delivery models and shown successful vascularization of the biomaterial, demonstrating the versatile vascular regenerative capabilities of the hydrogel design.
Abstract P2056: A Micropeptide Encoded By An Endogenous Retroviral-derived Lncrna Regulates Cardiomyocyte Multinucleation
Authors: Kelly M Anderson, Pornthida Poosala, Sean Lindley, Douglas M Anderson
Journal: Circulation Research (2022)

Endogenous retroelements (REs) are pervasive in mammalian genomes, and consist mostly of silenced mobile genetic sequences long considered to be ‘junk DNA.’ In a few known instances, mammalian genomes have captured and repurposed endogenous retroviral sequences to play important roles in regulating mammalian cell functions, notably cell fusion. Our previous work has shown that some mammalian transcripts misannotated as lncRNAs can be translated into small functional peptides, called micropeptides. Here we report the discovery of a cardiac-specific micropeptide encoded by a misannotated lncRNA transcribed from a murine endogenous retroviral element. We named this transcript CARDI, and the encoded micropeptide CARDI-B, since translation of the micropeptide occurs from the second ATG start codon. We show that CARDI-B shares homology with a singular domain of a retroviral protein and localizes to the nucleus. Genetic deletion of CARDI-B in mice from an isogenic background significantly increased the frequency of binucleated cardiomyocytes, at the expense of mononucleated cardiomyocytes. In line with previous reports that the frequency of mononucleated cardiomyocytes is correlated with cardiac regeneration, we show that CARDI-B-deficient hearts perform worse following myocardial infarction injury. These data reveal that functional micropeptides can be concealed within mammalian retroelements, and suggests that RE-derived micropeptides may contribute to the natural variation in heart development and regeneration observed across species.
Abstract P3041: DWORF Overexpression In The Heart Results In Enhanced Mitochondrial Function
Authors: Omar Brito-Estrada, Aaron Gibson, Cat Makarewich
Journal: Circulation Research (2022)

Background: Calcium (Ca 2+ ) plays a pivotal role in regulating physiological processes in the heart including excitation-contraction coupling, Ca 2+ signaling, and mitochondrial function. It is well established that both Ca 2+ dysregulation and altered cardiac energy metabolism are universal features of heart failure (HF). Dwarf open reading frame (DWORF), a newly discovered microprotein, increases SERCA2a activity in part by relieving SERCA2a inhibition by phospholamban (PLN). DWORF overexpression via cardiac specific transgene or gene therapy is cardioprotective in genetic and experimental HF mouse models. In addition to activation of SERCA2a by PLN displacement, preliminary data indicates that enhanced mitochondrial function contributes to the cardioprotective effects of DWORF. Here we tested the effects of DWORF overexpression on cardiac mitochondrial function and Ca 2+ dynamics. Methods and Results: We isolated mitochondria from wildtype (WT) and DWORF transgenic (Tg) mouse hearts and measured O 2 consumption rates using a Seahorse mito stress test. We found that DWORF Tg mice have increased respiratory capacity compared to WT (DWORF Tg 1150.0 pmol/min, WT 799.4 pmol/min +/-146.2) and enhanced spare respiratory capacity (DWORF Tg 489.1 pmol/min, WT 229.4 pmol/min +/- 102.1). To measure mitochondrial Ca 2+ dynamics, we performed Ca 2+ retention capacity assays and found that DWORF Tg mice have increased Ca 2+ uptake rates compared to WT (DWORF Tg 32.32 s, WT 46.46 s +/- 4.39). Western blot analysis of the mitochondrial Ca 2+ uniporter (MCU) in mito extracts from WT and DWORF Tg hearts indicated elevated levels of MCU. However, electron transport chain (ETC) protein expression indicated no significant differences, indicating the enhanced mitochondrial activity observed in DWORF Tg mice was not due to increased ETC levels. Conclusions: DWORF Tg mouse heart mitochondria exhibit increased maximal respiration rates and enhanced spare respiratory capacity. However, ETC protein levels are unchanged, indicating altered enzyme activity rather than expression. Additionally, mitochondria from DWORF Tg hearts uptake Ca 2+ faster into the mitochondria through the increased levels of MCU, suggesting enhanced mitochondrial Ca 2+ dynamics.
Abstract P2110: Myosin-7 E848G Mutation Reduces Contractility And Activates P53-associated Intrinsic Apoptosis In Patient-derived Induced Pluripotent Stem Cell Model Of Hypertrophic Cardiomyopathy
Authors: Alexander Loiben, Clayton Friedman, Wei-Ming Chen, Leslie Chao, Kai Chun Yang
Journal: Circulation Research (2022)

Introduction: Familial hypertrophic cardiomyopathy (HCM), affecting 1 in 500 adults, is characterized by idiopathic thickening of the heart and impaired systolic function. Mechanisms through which cardiac sarcomeric mutations manifest in HCM are poorly understood. Hypothesis: We previously identified a novel MYH7 E848G mutation associated with HCM that manifested as left ventricular hypertrophy, diffuse fibrosis, and severe systolic dysfunction. We hypothesize that the E848G variant impairs tissue contractility and causes cell death in a dose-dependent manner. Methods: We created MYH7-EGFP expressing cardiomyocytes (CMs) with MYH7 wt/wt-EGFP , MYH7 wt/E848G-EGFP , or MYH7 E848G/E848G-EGFP alleles using patient-derived induced pluripotent stem cells (hiPSCs), CRISPR-Cas9 gene-editing, and standard differentiation protocols. CMs were cocultured with stromal cells to create 3D engineered heart tissues (EHTs) or cultured as a monolayer in high calcium stress media. EHTs were enzymatically digested and GFP+ cardios were sorted for analysis. Results: Monolayer MYH7 wt/E848G-EGFP CMs had higher TUNEL + percentage (13.1 ± 2.1%) at d33 and lower cell survival (52.0 ± 6.6% of d30 seeded) at d42 relative to MYH7 wt/wt-EGFP (6.6 ± 2.6%; 107 ± 2.1%). Screening revealed elevated levels of proteins associated with intrinsic apoptosis (p53, 1.93 ± .03-fold; p21, 1.66 ± .10-fold; Bax, 1.56 ± .11-fold) in MYH7 wt/E848G-EGFP CMs at d35 relative to MYH7 wt/wt-EGFP . MYH7 wt/E848G-EGFP (n = 20; 67.5 ± 9.0 μN) had reduced maximum active twitch force relative to MYH7 wt/wt-EGFP (n = 19; 226 ± 17 μN) 14 days post-cast. MYH7 wt/E848G-EGFP EHTs had lower percentage of EGFP + cells (13.5 ± 0.8%) 7 days post-cast relative to MYH7 wt/wt-EGFP (25.8 ± 0.8%). EGFP + cells sorted from 7 day post-cast MYH7 wt/E848G-EGFP EHTs had elevated TP53 transcription (1.67 ± .29-fold) relative to MYH7 wt/wt-EGFP . Conclusion: These results suggest MYH7 E848G mutation reduces sarcomere contractility and induces p53-associated cell death. High calcium stress media induces cell death that likely contributes to contractile dysfunction seen in MYH7 E848G HCM. Ongoing studies will elucidate mechanisms through which E848G activates intrinsic apoptosis.